Display Settings:

Format

Send to:

Choose Destination
Methods Enzymol. 2010;470:695-707. doi: 10.1016/S0076-6879(10)70029-X. Epub 2010 Mar 1.

Overexpression and purification of integral membrane proteins in yeast.

Author information

  • 1Department of Biochemistry and Biophysics, University of California at San Francisco, San Francisco, California, USA.

Abstract

The budding yeast Saccharomyces cerevisiae is a viable system for the overexpression and functional analysis of eukaryotic integral membrane proteins (IMPs). In this chapter we describe a general protocol for the initial cloning, transformation, overexpression, and subsequent purification of a putative IMP and discuss critical optimization steps and approaches. Since expression and purification are often the two predominant hurdles one will face in studying this difficult class of biological macromolecules the intent is to outline the general workflow while providing insights based upon our collective experience. These insights should facilitate tailoring of the outlined protocol to individual IMPs and expression or purification routines.

Copyright © 2010 Elsevier Inc. All rights reserved.

PMID:
20946832
[PubMed - indexed for MEDLINE]

LinkOut - more resources

Full Text Sources

Other Literature Sources

Molecular Biology Databases

PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for Elsevier Science
    Loading ...
    Write to the Help Desk